Effects Of Different Impact Factors On The Hairy Root Growth And Its Secondary Metabolites Of Pharbitis Purpurea

Pharbitis purpurea (L.) Voigt is annual, family convolvulaceae herb. Seeds can be used as medicine and it tastes bitter, quality of cold, toxic, in the lungs, kidney and large intestine. Has the discharge water aperient, sputum polyester away to drink, and insecticidal attack product function. So it is widely used in the treatment of water swelling full, obstruction of urination and defecation, retention of phlegm and fluid, cough result from breathing reversal and enterozoic abdominalgia with the function of water release and catharsis, phlegm and carouse, insecticidal and elimination stasis.Growth characteristics of the hairy root of Pharbitis purpurea and the contents of coffee acid, ethyl caffeate by multiple proliferation method and high-performance liquid chromatography (HPLC), the objective of the present study was to know effect the different impact factor of culture medium type, inoculation amount, type of carbon source, concentration, temperature, pH value, hormone, inducible factor on growth characteristics of the hairy root and the contents accumulation of coffee acid, ethyl caffeate.Hairy root growth and active ingredients accumulation of coffee acid, ethyl caffeate were affected by the type of culture medium, inoculation amount, type of carbon source, concentration, pH value, temperature in the culture of Pharbitis purpurea. The results showed that the optimum culture condition was1/2MS fluid culture medium with3%sucrose concentration, pH value ranged from5.5to6.0,0.1g inoculation amount and the temperature was27±2℃.2,4-D,6-BA, NAA, KT, GA3could affect hairy root growth and accumulation of coffee acid, ethyl caffeate, but the growth of biomass and contents of coffee acid, ethyl caffeate increase were not synchronous during the research process.2,4-D could inhibit growth of fairy root of Pharbitis purpurea, high concentration2,4-D could significantly promote coffee acid, ethyl caffeate synthesize, but the contents of coffee acid, ethyl caffeate production gradually reduced;6-BA slight inhibit the growth of the hairy root, and no significant influence the contents of hairy root dry weight and coffee acid, ethyl caffeate; GA3did not have significant effect of hairy root growth, low concentration of GA3had simulative effect of coffee acid, ethyl caffeate,0.5mg/L GA3the contents of coffee acid, ethyl caffeate was the highest, and promoting effect was strong. When the concentration of GA3continues to increase, the contents of coffee acid, ethyl caffeate induce gradually; NAA, KT could accelerate fairy root growth and the contents of coffee acid, ethyl caffeate. NAA the best concentration was0.5mg/L, KR the best concentration was1.0mg/L～1.5mg/L.Respectively using different concentrations of MJA, SA, Aspergillus niger to deal with fairy root of Pharbitis purpurea. The results showed that different inducible factors had different effects of fairy root of Pharbitis purpurea. The concentration of MJA and Aspergillus niger were the higher in the culture medium could inhibit, SA did not have effect of fairy root. Different concentration of MJA accelerated synthesis of coffee acid, ethyl caffeate, the higher the concentration, the stronger the accelerating function. When MJA concentration was100μ mol/L, coffee acid production was the highest, was330.6μg, was1.12times of no inducible factors, concentration of MJA was50μ, mol/L the contents of ethyl caffeate was highest, was42.57μg, was1.07times of no inducible factors. Concentration of SA was30μmol/L, the contents of coffee acid, ethyl caffeate was highest, were0.0286%,0.0034%. The contentss of coffee acid, ethyl caffeate were454.74μg,69.96μg. The concentration of Aspergillus niger was40mg/L, the contentss of coffee acid, ethyl caffeate were the highest, were454.74μg、69.96μg respectively. Thing of biomass liveweight of fairy root and the contents of coffee acid, ethyl caffeate, when the concentration of MJA was50～100μmol/L or the concentration of SA was30μmol/L or the concentration of Aspergillus niger was40mg/L, were appropriate for growth of fairy root and the contentss of coffee acid, ethyl caffeate were the highest.The results showed that the contents of coffee acid, ethyl caffeate were affected by different hormone combination, different inducible factor combination, hormone and inducible factor combination to fairy root of Pharbitis purpurea. The best combination culture medium were0.5mg/L NAA and1.0mg/L KT,100μmol/L MJA and30μmol/L SA,0.5mg/LNAA and30p. mol/L SA combination. The contents of coffee acid were376.2μg,387.2μg,452.1μg and ethyl caffeate were49.8μg,48.63μg,51.2μg.